The present invention relates to an antibody to a peptide including at least 8 successive amino acids of an amino acid sequence represented by formula [1] described below, and relates to hybridomas, methods for producing the same and uses thereof.
Many cell growth factors have been isolated and their structures have been elucidated since the discovery of epidermal growth factor (hereinafter referred to as EGF) and nerve growth factor (hereinafter referred to as NGF).
Cell growth factors are useful for the elucidation of cell differentiation mechanism and cell proliferation mechanism, and some of them, including human EGF, are expected to be useful as medicines. Therefore, studies thereon have become increasingly prevalent in recent years.
As to human NGF, its genomic DNA has been isolated, but has not been expressed in a host cell. Hence, the investigations of producing human NGF in large amounts and of using it have not been advanced.
The present inventors cloned a cDNA sequence coding for a polypeptide (I) which showed about 60% homology to human NGF from human glioma cDNA libraries (Japanese Patent Unexamined Publication Mo. 1-193654/1989 which corresponds to EP Application No. 90 104 419.8 and U.S. patent application Ser. No. 07/488,696, refer to FIGS. 1 to 4). The polypeptide (I) includes the following amino acid sequence represented by formula [1] in a molecule thereof: EQU TyrAlaGluHisLysSerHisArgGlyGluTyrSerValCys AspSerGluSerLeuTrpValThrAspLysSerSerAlaIle AspIleArgGlyHisGlnValThrValLeuGlyGluIleLys ThrGlyAsnSerProValLysGlnTyrPheTyrGluThrArg CysLysGluAlaArgProValLysAsnGlyCysArgGlyIle AspAspLysHisTrpAsnSerGlnCysLysThrSerGlnThr TyrValArgAlaLeuThrSerGluAsnAsnLysLeuValGly TrpArgTrpIleArgIleAspThrSerCysValCysAlaLeu SerArgLysIleGlyArg[1]
The polypeptide (I) is considered to have an action similar to that of NGF and important roles in vivo such as enhancement of differentiation, growth and proliferation of animal cells, enhanced expression of genes and induction of proteins and enzymes. This polypeptide has therefore the high possibility that it can be used as medicines.
Basic information as to the polypeptide (I) such as the distribution thereof in vivo, the production mode thereof or the mechanism of activity expression favors the development of the polypeptide (I) as medicines.
It is also important to know the amount of the polypeptide (I) exactly, when this protein is purified from a gene recombinant.
Previously, the amount of NGF has been calculated by assaying the neurite outgrowth action to PC12 cells. Also, the neurite outgrowth action to avian dorsal root ganglia has been utilized to calculate the amount of NGF. However, these assays have the disadvantages that the elaborate procedure is required and the measurement error is large, in addition, that a long time is required to obtain the result.
For these reasons, it has been desired to develop means of assaying the polypeptide (I) simply and exactly.